Serafini 2015 Abstract MiPschool London 2015
|Mitochondria-ER tethers: Identification by a FRET based high throughput screen.|
Event: MiPschool London 2015
The existence of points of close juxtaposition between mitochondria and endoplasmic reticulum (ER) [1,2] participates in lipid and Ca2+ homeostasis, for example by the generation of microdomains of high [Ca2+] on the mitochondrial surface  essentials for its exchange ion between the two organelles. Mitofusin2 (Mfn2) was the first recognized mitochondria-ER structural tether [1,2], and although some other proteins have already been described to be involved in the tethering between the two organelles [1,2], the molecular identity of these tethers mostly remains unknown.
In order to identify proteins that compose/modulate the juxtaposition between the two organelles, with or independently from Mitofusin2, we set up a genome wide screening based on modified inducible FRET based, mito-ER tethers probe (FEML) whereby the variations of the proximity between the two organelles are mirrored by changes in the FRET ratio value.
When FEML was transiently transfected in WT and Mfn2-/- MEFs, the two fusion proteins correctly localize respectively in mitochondria and ER, and, as expected, a change in the FRET ratio signal has been observed upon stimulation with Rapamycin.
Our genome wide, high content imaging approach will be instrumental to recognize both the negative and positive regulators of the mitochondria-ER contacts.
Labels: MiParea: mt-Structure;fission;fusion
Dept Biol, Univ Padova Dulbecco-Telethon Inst, Venetian Inst Mol Med, Padova, Italy. - [email protected]
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